ICS 77.100.40  
 
Reference number  
DRS 454: 2021  
© RSB 2021  
  
 RWANDA 
STANDARD  DRS 
454 
 
First edition  
Approved by RSB 2021 -mm-dd 
 
Alcohol -based hand sanitizer s — 
Specification  
 
   
DRS 454: 2021  
©RSB 2021  - All rights reserved                  ii 
 In order to match with technological development and to keep continuous progress in industries, 
standards are subject to periodic review. Users shall ascertain that they are in possession of the latest 
edition  
 
 
 
 
 
 
 
 
© RSB 2021 
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized 
in any form or by any means, electronic or mechanical, including photocopying and microfilm, without 
prior written permission from R SB. 
Requests for permission to reprodu ce this document should be addressed to : 
Rwanda Standards  Board  
P.O Box 7099 Kigali -Rwanda  
KK 15 Rd, 49  
Tel. +250 788303492  
Toll Free: 3250  
E-mail: info@rsb.gov.rw  
Website: www.rsb.gov.rw  
ePortal: www.portal.rsb.gov.rw   DRS 454: 2021  
iii                ©RSB 2021  - All rights reserved  
 Contents  Page 
1 Scope  ................................ ................................ ................................ ................................ .............  1 
2 Normative references  ................................ ................................ ................................ ...................  1 
3 Terms and definitions  ................................ ................................ ................................ ..................  1 
4 Requirements  ................................ ................................ ................................ ................................  2 
4.1 General requirements  ................................ ................................ ................................ ..................  2 
4.2 Specific requirements  ................................ ................................ ................................ ..................  2 
5 Packaging and labelling ................................ ................................ ................................ ...............  2 
5.1 Pack aging  ................................ ................................ ................................ ................................ ...... 2 
5.2 Labelling  ................................ ................................ ................................ ................................ ........  3 
6 Sampling  ................................ ................................ ................................ ................................ ....... 4 
6.1 General  ................................ ................................ ................................ ................................ ..........  4 
6.2 Sample for inspection  ................................ ................................ ................................ ..................  4 
6.3 Sample for testing  ................................ ................................ ................................ ........................  4 
Annex  A (normative)  Determination of antimicrobial efficacy ................................ .............................  5 
A.1 Outline of the method  ................................ ................................ ................................ ..................  5 
A.1 Apparatus  ................................ ................................ ................................ ................................ ...... 5 
A.2 Media ................................ ................................ ................................ ................................ ..............  6 
A.3 Selection of the most resistant organism by the minimum inhibitory concentration test  ... 7 
A.4 Preparation of inoculum  ................................ ................................ ................................ ..............  8 
A.5 Preparation of the sanitizer dilutions  ................................ ................................ .........................  8 
A.6 Test procedure  ................................ ................................ ................................ ..............................  8 
A.7 Interpretation of results  ................................ ................................ ................................ ...............  9 
Annex  B (normative)  Determination of dermal irritation  ................................ ................................ .... 10 
B.1 Test panel  ................................ ................................ ................................ ................................ .... 10 
B.2 Procedure  ................................ ................................ ................................ ................................ .... 10 
Annex  C (normative)  Freedom from visible impurities  ................................ ................................ ...... 11 
 DRS 454: 2021  
©RSB 2021  - All rights reserved                  iv 
 Foreword  
Rwanda Standard s are prepared  by Technical Committees and approved by R wanda Standards Board (RSB) 
Board  of Directors in  accordance with the procedures of RSB, in compliance with Annex 3 of the WTO/TBT 
agreement on the preparation, adoption and application of standards.  
The main task of technical committees is to prepare national standards.  Final Draft Rwanda S tandards adopted 
by Technical committ ees are ratified  by me mbers of RSB Board of Director s for publication and gazettment as 
Rwanda Standards.  
DRS 454 was prepared by Technical Committee RSB/TC 024, Organic and Inorganic Chemicals . 
In the preparation of thi s standard, reference was made to the following standard s: 
1) RS EAS 789: Instant hand sanitizers — Specification  
2) FDARS 1470: Hand sanitizer (alcohol -based) — Specification  
The assistance de rived from the above source is hereby acknowledged with thanks.  
Committee membership  
The following organizations were represented on the Technical Committee on Organic and Inorganic Chemicals 
(RSB/TC 024) in the preparation of this standard.  
Star Construction and Consultancy Ltd  
Rwanda Inspectorate, Competition and Con sumer Protection Authority  
Rwanda Food and Drugs Authority  
Rwanda Investigation Bureau  
Rwanda Forensic Laboratory  
Rwanda Social Security Board  
Rwanda Environment Management Authority  
BARANYUZWE Cosmetics Ltd  
SULFO Rwanda Industries Ltd  
UBURANGA Natural Pro ducts  DRS 454: 2021  
v                ©RSB 2021  - All rights reserved  
 Divine Hope Company Ltd  
University of Rwanda/College of Medicine and Health Sciences  
University of Rwanda/College of Sciences and Technology  
University of Rwanda/College of Education  
Rwanda Polytechnic – IPRC Kigali  
Standards for Sustainability (SfS)  
Rwanda  Standards Board  (RSB) – Secretariat  DRS 454: 2021  
©RSB 2021  - All rights reserved                  vi 
 Introduction  
The alcohol contained within hand sanitizers, when rubbed on the surface of skin is effective in killing 99.9% of 
dangerous germs on the skin. The type of alcohol used in most hand sanitizers is eth yl alcohol. Ethyl alcohol is 
the active ingredient in most hand sanitizers. A concentration of 60% to 95% alcohol in hand sanitizing product 
is recommended. However, different scientific literatures have shown that alcohol, at concentration of above 
70% is  a potential virucidal agent inactivating all of the lipophilic viruses and many hydrophilic viruses (e.g., 
adenovirus, enterovirus, rhinovirus and ratoviruses).  
There are also several other non -active ingredients in hand sanitizer, the second most concent rated ingredient 
is water.  
 
 DRS 454 : 2021  
1                ©RSB 2021  - All rights reserved  
 Alcohol -based hand sanitizers  — Specification  
1 Scope   
This Draft Rwanda Standard specifies the requirements, sampling and test methods for alcohol -based hand 
sanitizers.  
2 Normative references   
The following documents are referred to in the text in such a way that some or all of  their content constitutes 
requirements of this document.  For dated references, only the edition  cited applies. For undated references, the 
latest edition of the referenced document (including any amendments) applies.  
RS EAS 377, Cosmetic Products (all parts)  
AOAC 942.06, Alcohol by Volume in Distilled Liquors Pycn ometer Method  
RS ISO 10523, Water quality — Determination of pH  
 
3 Terms and definitions   
For the purposes of this standard , the follow ing terms and definitions apply.  
3.1 
hand sanitizer  
antiseptic agents used to cleanse the hands with the aim to protect and  prevent the passage of bacteria, virus 
and other pathogens that can cause infections.  
3.2 
alcohol  
ethanol (ethyl alcohol, C 2H5OH), isopropanol (isopropyl alcohol, CH 3CHOHCH 3), n-propanol (1 -propanol, 
CH 3CH 2CH 2OH) or the mixture of them.  
3.3 
antimicrobial  efficacy 
efficiency of the product to kill or reduce microorganisms, such as bacteria, fungi and viruses.   
©RSB 2021  - All rights reserved                  2 
 3.4 
dermal irritation  
production of reversible damage of the skin following the application of a test substance for up to 4 hours.  
4 Requirements  
4.1 Genera l requirements  
4.1.1  The product shall be in the form of liquid or gel.  
4.1.2  The product shall be clear and free from visible impurities.  
4.1.3  The product shall not have any disagreeable odour or smell. It may be coloured or not.  
4.1.4  The substances use d in the formulation shall conform to all parts of RS EAS 377.  
4.2 Specific requirements  
The product shall also comply with the specific requirements given in the table 1 when tested in accordance 
with the corresponding test method.  
Table 1 — Specific requirem ents for alcohol -based hand sanitizers  
S/N Characteristic Requirement Test method 
i) Alcohol content (ethanol and/or 
isopropanol, n-propanol), % v/v, min. a 60.0 
 AOAC 942.06  
ii) pH, neat 6 – 8 RS ISO 10523  
iii) Antimicrobial  efficacy To pass test Annex A 
iv) Dermal irritation  To pass test  Annex B  
v) Physical impurities  To pass test  Annex C  
a The product that has been declared as potential virucidal agent shall contain more than 70% of 
alcohol content in order to be effective as a  virucidal agent inactivating all of the lipophilic viruses 
and many hydrophilic viruses (e.g., adenovirus, enterovirus, rhinovirus and ratoviruses)  
5 Packaging and labelling  
5.1 Packaging  
5.1.1  The packaging shall ensure integrity of the product during handling, storage and tr ansportation.  
5.1.2  Bulk packaging: Only the sanitizer of the same type and the same batch shall be packed together in 
one bulk package  
5.1.3  The closure shall not be made of cork or of any other material that contains cork.  DRS 454 : 2021  
3                ©RSB 2021  - All rights reserved  
 5.2 Labelling  
The following informa tion shall appear in legible and indelible labelling on each container or on a label securely 
attached to each container:  
a) name of the product as “alcohol -based hand sanitizer”;  
b) name and full address of the manufacturer;  
NOTE  The name, physical address of t he distributor/supplier may be added as required.  
c) percentage of alcohol used  
NOTE  The tolerance limit from the declared alcohol content shall be less than 1  
d) list of ingredients in descending order.  
e) net content  
f) batch identification;  
g) manufacture and expiry dates;  
h) general instructions for use;  
i) storage conditions;  
j) the following cautionary warnings:  
1) “do not allow the sanitizer to come into contact with eyes”;  
2) “keep out reach of children”;  
3) “if swallowed contact a doctor”; and  
4) “highly flammable, keep away from fire or flame”.   
©RSB 2021  - All rights reserved                  4 
 6 Sampling  
6.1 General  
The following sampling procedure shall be applied in determining whether a lot submitted for inspection and 
testing complies with the relevant requirements of this standard. The sample so drawn shall be deemed to 
represent the lot.  
6.2 Sample for inspection  
After inspecting the lot for compliance with Clause 4, take, at random, the number of containers, as relevant, 
shown in column 2 of Table 2, relative to the appropriate lot size shown in column 1.  
Table 2 — Samples for inspec tion and testing  
Lot size 
(num ber of containers) Sample size for ph ysical 
examination 
(num ber of containers) Sample size for micro biological 
examin ation 
(num ber of containers) 
0 to 5 000  3 3 
5 001  to 12 5 00 6 3 
12 501 to 25 0 00 9 3 
25 001 to 50 0 00 16 3 
50 001 upwards 30 3 
6.3 Sample for testing  
After inspection of the containers taken in accordance with 6.2,  
a) take, at random, half the number of containers and use them for the storage stability test; and  
b) thoroughly mix the contents of each of the remaining  containers and, take from each container the lesser 
of the total volume and 250 mL, and obtain a composite test sample by combining and thoroughly mixing 
these quantities. Use these samples for testing for compliance with the requirements of Clause 4.  
 DRS 454 : 2021  
5                ©RSB 2021  - All rights reserved  
 Annex  A 
(normative)  
 
Determination of antimicrobial efficacy  
A.1 Outline of the method  
A.1.1  The sanitizer is tested at the recommended ‘use -dilution’ and concurrently at 0.5 and 1.5 times that 
dilution. The test consists of challenging the diluted sanitizer with bacterial inoculum, withdrawing a sample after 
a given time and culturing the sample in a suitable recovery medium. After this sampling, the mixture is again 
challenged by a second inoculum and after a second interval, is again sampled for culturing. This process is 
then repeated to provide a third challenge.  
A.1.2  The sample is considered to have passed or failed the test according to the extent of growth shown in 
the first two cultured samples . 
A.1 Apparatus  
A.2.1  Facility , for incubation at 37 ± 1 °C.  
A.2.2  Facility, for incubation at 27 ± 1 °C.  
A.2.3  Stop clock, indicating in seconds.  
A.2.4  Facility, for refrigeration at 4 ± 1°C.  
A.2.5  Universal containers ― Made of glass and having metal tops with rubber liners. Plastic containers or 
glass containers wi th plastic tops shall not be used.  
A.2.6  Test tubes — 19 mm X 150 mm.  
A.2.7  Filter paper, No. 4 whatman (sterile) or equivalent.  
A.2.8  Facility, for autoclaving at 121 ± 1°C.  
A.2.9  Pipette, capable of dispensing 0.02 ± 1°C 0.005 ml.  
A.2.10  pH meter  
A.2.11 Facility, to sterilize by filtration.  
A.2.12  150 μm test sieve.  
A.2.13  Oven, capable of maintaining temperature at 100 ± 1 °C.   
©RSB 2021  - All rights reserved                  6 
 A.2 Media  
A.3.1  Growth media for test organisms. Wright and Mundy Broth with Dextrose (WMBD).  
A.3.1.1  Dispense 10 ml and 6 m l quantities of the Wright and Mundy Broth into universal bottles, and autoclave 
at 121 ± 1 °C for 12 minutes.  
A.3.1.2  Add to this medium, 10 per cent (m/V) dextrose solution sterilized by filtration, to give a final dextrose 
concentration of 0.1 per cent (m/v), (i.e. to 10 mL broth add 0.1 dextrose solution and to 6.0 mL broth add 0.06 
mL dextrose solution).  
A.3.2  Recovery medium ― A nutrient broth prepared as follows:   
A.3.2.1  Composition   
 Beef extract 10 g  
 Peptone 10 g  
 Sodium chloride 5 g  
 Polyoxyethylene sorbitan mono -oleate 30 g  
A.3.2.2  Preparation ― Add the ingredients to 1000 mL of water. Mix well. Dispense 10 ml quantities into test 
tubes and autoclave at 121 ± 1°C for 15 minutes . 
A.3.3 Hard water — Standard hard water with 342 mg/L (p pm) hardness prepared as follows:  
Dissolve 0.304 g of anhydrous calcium chloride hexahydrate (MgCl2.6H 2O) in distilled water and make up the 
volume to one litre. Sterilize the standard hard water by autoclaving at 121 ± 1° C for 15 minutes. Allow this to 
reach room temperature before use . 
A.3.4 Yeast suspension  
A.3.4.1  Weigh to the nearest gram about 65 g of active dry yeast. Cream by the gradual addition of sterile hard 
water (A.3.3) using a heavy glass rod for stirring. Decant the creamed portion into a  flask, add more hard water 
to any lumpy residue remaining and repeat the creaming and decantation until no residue remains, and 500 ml 
of hard water has been used.  
A.3.4.2  Shake the contents of the flask vigorously and strain -through a 150 m sieve (A.2. 12) breaking down 
any remaining lumps.   
A.3.4.3  Add 500 mL sterile hard water, shake vigorously.   
A.3.4.4  Transfer 50 ml or 100 mL portions into screw -capped bottles, screw the caps tightly and autoclave at 
121 ± 1°C for 15 minutes. Allow the autoclave to cool without releasing the pressure. Store cold but not freezing.   DRS 454 : 2021  
7                ©RSB 2021  - All rights reserved  
 A.3.4.5  Dry two glass petri -dishes to constant mass. Into each of these dishes, pipette 25 mL of sterilized yeast 
suspension and dry to constant mass at 100°C. Calculate the average solids content of the suspension.   
A.3.4.6  Before use, pipette 25 ml of the sterilized yeast suspension into a beaker. Determine the pH using a 
glass electrode, and determine the volume of 40 g/L sodium hydroxide solution needed to adjust the pH to 7.0 
± 0.1.   
A.3.4.7  Immediately before use, add to each bottle of sterilized yeast suspension a volume of sterile hard water 
and a volume of 40 g/l sodium hydroxide calculated to adjust the concentration of dry yeast to 5 per cent (m/V) 
and the pH to 7.0 ± 0.1. Discar d prepared yeast, two weeks after preparation.  
A.3.5  Ringers solution, 25 per cent (V/V)  
Dissolve 9.00 g of sodium chloride, 0.42 g of potassium chloride, 0.24 g of anhydrous calcium chloride and 0.20 
g of sodium bicarbonate in water and dilute to 1000 m l. Add 1 volume of this solution to 3 volumes of water to 
give a 25 per cent solution. Dispense into test tubes fitted with suitable closures and sterilized by auto -claving 
at 121 ± 1°C for 15 minutes . 
A.3 Selection of the most resistant organism by the minimu m inhibitory 
concentration test  
A.4.1  The following organisms shall be used for the test:  
 Pseudomonas aeruginosa (NCTC 6749 or equivalent)  
 Proteus vulgaris (NCTC 4635 or equivalent)  
 Staphyloccus aureus (NCTC 4163 or equivalent)  
These organisms may be ob tained as freeze dried cultures. Once sub -cultured, the organisms shall be 
maintained on agar slopes of suitable nutrient medium at 4 ± 1°C.  
A.4.2  Subculture each organism daily into a universal bottle containing 6 ml of growth medium (A.3.1) and 
incubate  for 24 ± 2 h at 37 ± 1°C.  
A.4.3  Dilute one part of freshly grown sub -culture of each organism, which is at least a fifth sub -culture and 
not more than a fourteenth, with ten parts of the growth medium (A.3.1) before dilution, the P. aeruginosa, 
culture sh all be filtered using a whatman No.4 filter paper.  
A.4.4  Prepared three sets of ten, doubling dilutions of the sanitizer in universal containers (A.2.5). For this 
purpose dilute the neat sanitizer in the growth medium (A.3.1) or the recovery medium (A.3.2 ) to give a final 
volume of 5 ml of the diluted sanitizer for each dilution.  
A.4.5  Inoculate each dilution in one set with 0.02 mL of a diluted culture of one organism (see A.4.3).  
A.4.6  Incubate all the three sets of inoculate dilutions at 37 ± 1°C for 72 hours, and examine to determine 
the organism most resistant to the sanitizer, that is the organism for which the minimum inhibitory concentration 
is highest.   
©RSB 2021  - All rights reserved                  8 
 A.4 Preparation of inoculum  
A.5.1  Daily sub -cultures of the test organism selected as in A.4.6 shal l be grown in 6 ml quantities of the 
growth medium (A.3.1) and incubated at 37 ± 1°C for 24 ± 2 hours.  
A.5.2  The day before the test, inoculate 10 ml of the growth medium (A3.1) with the test organism from a 
daily sub -culture and not more than a fourteenth . Incubate the inoculated, broth at 37 ± 1°C for 24 ± 2 hours.  
A.5.3  Add 6 ml of the test organism culture (A.5.1) and (A.5.2) to 4 ml of the yeast suspension (A.3.4) thus 
making a final concentration of 2 per cent (m/V) of yeast in the yeast/organism susp ension. If a culture of P. 
aeruginosa is used, it shall be filtered using a whatman No.4 filter paper before addition . 
A.5.4  Shake the yeast/organism suspension for one minute with a few sterile glass beads. Immediately 
before the test, count the number of  viable organisms in the inoculum by decimal dilutions in 25 per cent Ringers 
solution (see A.3.5) and by the drop plate method. The viable count shall be not less than 108 organisms/ml or 
more than 1010 organisms/ml or the test results are considered inva lid. 
A.5 Preparation of the sanitizer dilutions  
Prepare three dilutions of the sanitizer in hard water (A.3.3) based on the recommended ‘use dilution’ of the 
sanitizer, as follows:  
A = 0.5 times the recommended ‘use -dilution’  
B = 1.0 times the recommended ‘use -dilution’  
C = 1.5 times the recommended ‘use -dilution’  
The sanitizer dilutions shall be prepared and tested on the same day.  
A.6 Test procedure  
A.7.1  The test shall be carried out at 27 ± 1°C.  
A.7.2  Dispense 3 mL of each dilution of sanitizer (A.6) into separ ate universal bottles labelled A, B, and C, 
then allow to equilibrate to 27 ± 1°C.  
A.7.3  Add 1 mL of the inoculum to A, B and C at 0, 1 and 5 minutes respectively and mix by swirling gently.  
A.7.4  Eight minutes after the addition of the inoculum, remove a sample of the inoculum/sanitizer mixture 
and put 0.02 ml into each of the first group of five tubes of recovery broths. Return the remainder of the mixture 
in the pipette to the universal container . 
A.7.5  Ten minutes after the first addition of the inoculum, add another 1 ml of the inoculum to each of the 
sanitizer dilutions and mix by swirling gently.  DRS 454 : 2021  
9                ©RSB 2021  - All rights reserved  
 A.7.6  Eight minutes later, remove a sample of the mixture as put before (A.7.4) and put 0.02 mL into each 
of the second group of five tubes of recover y broths.  
A.7.7  Twenty minutes after the first addition of the inoculum, add a further 1 mL of inoculum to each of the 
sanitizer dilutions and mix by swirling gently.  
A.7.8  Eight minutes later, remove a sample of the mixture as before and place 0.02 ml int o each of the third 
group of five tubes of recovery broths.  
A.7.9  Swirl the recovery broths and incubate at 37 ± 1°C for 48 ± 2 h. Examine the growth and record the 
results . 
A.7 Interpretation of results  
A.8.1  The sanitizer, shall be regarded as having passed the test at the recommended ‘use dilution’ if there 
is no growth in at least two of the five recovery broths for the first and second additions of the inoculum.  
A.8.2  To be acceptable, an instant hand sanitizer shall pass the test on three separate occasio ns using 
freshly prepared sanitizer and freshly prepared inoculum on each occasion . 
 
   
©RSB 2021  - All rights reserved                  10 
 Annex  B 
(normative)  
 
Determination of dermal irritation  
B.1 Test panel  
A test panel that consists of three men and three women, none of whom is known to have an abnormally 
sensitiv e skin or has an injury or abrasion on the hands.  
B.2 Procedure  
Place approximately 5 mL of the test sample onto the cupped palm of one hand of each member of the panel, 
and get him or her to spread the Hand Sanitizer over the back and between the fingers of t he other hand, and 
rub it thoroughly into the skin for 2 min. Repeat this procedure twice, with 30 min intervals between applications. 
Do not allow a treated hand to be washed until 2 h after the last application of the test sample.  
Immediately after the t ests, and again 2 h, 24 h and 48 h later, examine the treated hand of each member of 
the panel for any signs of irritation or inflammation, using the untreated hand as a control.  DRS 454 : 2021  
11                ©RSB 2021  - All rights reserved  
 Annex  C 
(normative)  
 
Freedom from visible impurities  
C.1 A sample of approximately 50 mL of each test sample is spread over the bottom of a 150 mm diameter 
Petri dish.  
C.2 When viewed at a range of approximately 600 mm, the number of visible specks of impurities shall not 
exceed five.  
 
  
©RSB 2021  - All rights reserved                  12 
 Bibliography  
[1] World Health Organization (WHO), Guidelines on Hand Hygiene in Health Care , 2009  
[2] World Health Organization (WHO), Guide to local production — WHO -recommended Handrub 
Formulations, 2009.  
[3] Rwanda Foods and Drugs Authority (FDA), Guidelines on Preparation of Alcoholic Based Hand 
sanitizers, 2020.  
 
 DRS 454: 2021  
 
Price based on …..pages  
©RSB 2021  - All right s reserved